Composite

Part:BBa_K5181015

Designed by: Mayukh Mitra   Group: iGEM24_IISER-TVM   (2024-09-04)


tet_RBS_P450-CPR_fusion protein (BC_linker)

This part is a bicistronic composite part designed by team IISER-TVM. It consists of genes encoding a fusion protein of Cytochrome P450 monooxygenase and Cytochrome P450 reductase from Santalum album with BC linker under the control of tet promoter. It is a composite part comprising of basic parts - Self-sufficient tet_RBS (BBa_K5181009) and P450-CPR_fusion protein (BC_linker): BBa_K5181008 - This part is an artificial fusion protein of CYP450_truncated and CPR_truncated with BC linker.

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We performed Gibson assembly to obtain this part from three fragments synthesized by GenScript. The following Agarose gel electrophoresis map shows the amplified tet_CPR_P450 insert after bridge PCR to confirm Gibson assembly. The polymerase chain reaction was tested at four temperatures, as marked in Lanes 1,2,3,4. Band of size 4151 bp corresponds to amplified insert.

Part Image

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 636
    Illegal PstI site found at 972
    Illegal PstI site found at 1650
    Illegal PstI site found at 3441
    Illegal PstI site found at 3720
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1125
    Illegal PstI site found at 972
    Illegal PstI site found at 1650
    Illegal PstI site found at 3441
    Illegal PstI site found at 3720
    Illegal NotI site found at 7
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 730
    Illegal BglII site found at 3104
    Illegal XhoI site found at 2061
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 636
    Illegal PstI site found at 972
    Illegal PstI site found at 1650
    Illegal PstI site found at 3441
    Illegal PstI site found at 3720
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 636
    Illegal PstI site found at 972
    Illegal PstI site found at 1650
    Illegal PstI site found at 3441
    Illegal PstI site found at 3720
    Illegal NgoMIV site found at 1101
  • 1000
    COMPATIBLE WITH RFC[1000]


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